[District psychiatry. Male nurses facing future psychiatry].
Explore the source record for details and available documents.
Community Psychiatry↗
Biomedical subjects
J Folkersen
Publications and source records attributed to J Folkersen.
Enhancing effect of autologous human erythrocytes on generation of C3 cleavage products beyond iC3b.
The in vitro formation of C3d and C3c in fresh normal human serum (NHS) after addition of five different activators of the complement (C) system was studied. Following C-activation in NHS (n = 53) by Sephadex G-200 beads, the conversion of C3 was found to proceed to iC3b with a variable but restricted generation of C3d. Similar results were obtained by use of heat-aggregated IgG, Escherichia coli, zymosan, and cobra venom factor. However, comparing the C3d concentration following activation in the presence and absence of autologous red blood cells (RBC) at 37 degrees C the generated C3d was found to be 2- to 3-fold higher in the presence of RBC after 30, 60, and 210 min. Preincubation of RBC with polyclonal anti-CR1 antibodies resulted in a dose-dependent reduction of the amount of C3d generated. C-activation induced by Sephadex G-200 beads, in the absence of RBC, generated iC3b without a significant production of C3d. After removal of the activator beads, addition of RBC resulted in a decrease of iC3b and a clear increase in the C3c and C3d concentration within 3 h. Western blotting analysis of the C3d produced in the presence of RBC showed that the molecular weight (36 kilodaltons) was similar to that of C3d formed in vivo.
Blotting, Western↗
Heterogeneity of C4d and C3d and their complex formation with serum albumin.
High molecular weight (Mr around 175 KD) forms of C4d and C3d as well as free C4d and C3d (Mr about 40 KD) were demonstrable in normal human serum (NHS). Following in vitro C activation in NHS by delta IgG, the 40 KD C4d component increased markedly. By immunofixation it was shown that the high molecular forms of C4d and C3d reacted with biotinylated anti-human albumin IgG, whereas the 40 KD-free C4d and C3d fragments did not. Furthermore, the incorporation of anti-albumin IgG in the first dimensional gel in crossed immunoelectrophoresis caused retention of the 175 KD C4d component but not of free C4d. The 175 KD C4d had a distinctly higher electrophoretic migration velocity (post-albumin region) than the 40 KD C4d fragment. The C3d-, C4d-serum albumin complexes could not be dissociated by reducing agents (DTT, mercaptoethanol), a non-ionic detergent, or exposure to high and low ionic strength.
Complement Activation↗
Immunohistochemical aspects of immunological cross-reaction and masking of epitopes for localization studies on pregnancy-associated plasma protein A.
The influence of antibody absorption procedures and proteolytic pre-treatment of formaldehyde-fixed placental tissue on the localization of pregnancy-associated plasma protein A by immunoperoxidase technique was examined. Apparently monospecific IgG fraction of the anti-plasma protein applied directly on fixed tissue resulted in staining of connective tissue and a thin apical rim of the syncytiotrophoblast. Further absorption of the antibody with foetal connective tissue abolished this staining reaction. Pre-treatment of the fixed placental tissue with trypsin prior to application of the antibody, which had been absorbed with connective tissue, resulted in staining within the cytoplasm of the syncytiotrophoblast exclusively. Identical staining was seen when this IgG preparation was used directly on frozen placental tissue. The results point to the importance of the specificity of the antibody preparations and of proteolytic unmasking of epitopes when fixed tissues are used for localization studies of pregnancy-associated plasma protein A by immunoperoxidase technique.
Epitopes↗
Complement activation, circulating protease inhibitors and pregnancy-associated proteins in severe pre-eclampsia.
Circulating protease inhibitors, pregnancy-associated proteins and the split product of complement factor 3 (C3d) were measured in 14 women with severe pre-eclampsia and their matched controls. Only the mean levels of antithrombin III were observed to be significantly lower in pre-eclampsia (P less than 0.02).
Antithrombin III↗
Comparison of different antibody preparations against pregnancy-associated plasma protein-A (PAPP-A) for use in localization and immunoassay studies.
Four antibody preparations against pregnancy-associated plasma protein (PAPP-A) were compared in order to find an explanation for the contradictory results published on tissue localization, clinical usefulness and biological function of PAPP-A. One of the preparations studied was a rabbit anti-PAPP-A antiserum which has been offered for general scientific use (Bischof et al. 1979). Only the IgG fraction of anti-PAPP-A antisera which appeared to be monospecific and had been further absorbed with fetal connective tissue gave specific uniform staining of the cytoplasm of the syncytiotrophoblast exclusively. Circulating PAPP-A could not be detected by RIA employing this IgG preparation in the non-pregnant state, or before 18 days after conception. Circulating PAPP-A could be detected in all seven pregnant women studied within 4 weeks after conception. Identical results were obtained with a commercially available IgG fraction against PAPP-A.
Animals↗
Characterization of human pregnancy zone protein. Comparison with human alpha 2-macroglobulin.
Native human pregnancy zone protein (PZP), a close homolog of alpha 2-macroglobulin (alpha 2M), can be obtained in approximately 20% yield from pooled late pregnancy plasma or serum by a combination of polyethylene glycol precipitation, euglobulin precipitation, DEAE-Sephacel chromatography, zinc-chelate affinity chromatography, and negative affinity chromatography on insolubilized antibodies against human serum proteins. Both proteins are similarly organized as disulfide-bridged dimers of 360 kDa containing 180-kDa subunits. These dimers constitute the proteinase-binding units of PZP, and in contrast to alpha 2M, they appear to be only loosely associated, indicating a subtle difference in the quaternary structure of these alpha-macroglobulins. The preparations contain functionally intact beta-cysteinyl-gamma-glutamyl thiol esters, located in the same nonapeptide sequence as found in alpha 2M, and form complexes with a variety of proteinases in which a large fraction of the proteinase is bound covalently. Proteinases bound to PZP are still active and poorly accessible to reaction with large inhibitors like alpha 1-proteinase inhibitor. The structural and functional features of PZP indicate that PZP and alpha 2M, although extremely similar, may have different yet overlapping sets of proteinases as targets. It is possible that PZP mainly controls the activity of cellular proteinases released under conditions of increased cellular turnover and that PZP could be the human equivalent to the acute phase alpha-macroglobulins known in other species.
Chemical Precipitation↗
The selection of antibodies with defined desorption properties from precipitated immune complexes for use in immunoadsorption procedures.
A general method for preparing immunosorbents with preselected antibody avidity is described. The method, which is a modification of a method described previously, also includes immunospecific purification of the ligand prior to coupling on the gel matrix. Polyclonal anti-alpha-1-fetoprotein antibodies in precipitated immune complexes were separated according to their avidity (low, intermediate and high) by dissociation with agents of increasing efficiency. After solid-phase coupling the antigen binding activity of the separated antibody preparations was examined according to recovery, capacity and binding strength. Antibodies of intermediate avidity derived from the immune complexes demonstrated optimal properties for preparative affinity chromatography.
Antibody Affinity↗
Antigenic determinants of pregnancy-associated alpha 2-glycoprotein and alpha 2-macroglobulin defined by poly- and monoclonal antibodies.
Human alpha 2-macroglobulin and pregnancy-associated alpha 2-glycoprotein (PA alpha 2G) share several physicochemical characteristics. By the use of unabsorbed or absorbed polyclonal antibodies to these antigens, the existence of common epitopes in these molecules were demonstrated in crossed immunoelectrophoresis. Two monoclonal antibodies out of 9 raised against purified PA alpha 2G were demonstrated to react with both antigens, indicating close immunochemical relatedness between these macroglobulins. The findings might have functional implications.
Animals↗
Immunoelectrophoretic analysis of C4 split products expressing D but not C epitopes: influence of storage, Ca2+ and Ca2+-chelating agents.
Based on immunoelectrophoretic methods a heterogeneity in the electrophoretic mobility of C4d was observed. C4d was defined immunochemically as C4 molecules expressing D but lacking C epitopes. A beta-mobile form was observed when EDTA or heparin was not added to the sample prior to electrophoretic analysis. This component was generated during electrophoresis. Another C4d component migrating to the post-albumin region probably represented an in vivo generated split product. However, this C4d form was also produced during storage of serum or plasma at room temperature and its formation was enhanced in the presence of EDTA. Based on these findings standard conditions for collection and storage of clinical samples for quantification of C4d by electroimmunoassay are suggested.
Calcium↗
Specific assay for pregnancy-specific-beta 1-glycoprotein (Sp1-beta). Preparation of antiserum specific for SP1-beta by absorption with a crossreacting high molecular weight serum protein.
Two pregnancy-associated proteins reacting with antibody against pregnancy-specific beta 1-glycoprotein (PS beta G or SP1) were separated by means of ammonium sulphate precipitation, size chromatography and preparative zone electrophoresis. The antibody preparation was made monospecific to the beta-mobile protein by liquid phase cross-absorption of anti-SP1 immunoglobulin preparation with the isolated alpha2-mobile high molecular weight protein. The specificity of the absorbed antibody was tested in line immunoelectrophoresis, rocket immunoelectrophoresis and crossed immunoelectrophoresis.
Absorption↗
Quantification of the pregnancy-specific beta1-glycoprotein (SP1) by immunoprecipitation techniques: the influence of a cross-reacting high molecular weight alpha2-protein.
Two serum proteins with SP1 determinants were separated by means of size chromatography and preparative zone electrophoresis. One of the proteins had beta1 electrophoretic mobility and an estimated molecular weight of 90,000, and the other had alpha2-mobility and a molecular weight around 400,000. When available antibody preparations to SP1 were used, the high molecular weight protein formed immunoprecipitates only when polyethyleneglycol (PEG) was added to the agarose. It is shown that the high molecular weight protein has a pronounced effect on quantification of SP1 by rocket immunoelectrophoresis, whereas the effect was less pronounced but still considerable when quantification was performed by radial immunodiffusion.
Antigens↗
Demonstration of an analogue to human pregnancy zone protein in the African green monkey.
A serum protein in the grivet (Cercopithecus aethiops), immunologically cross-reacting with human pregnancy zone protein (PZP), is found in males as well as in nonpregnant females. The electrophoretic mobility, antigenic cross-reactivity, molecular weight, and carbohydrate structure appear analogous to PZP; however, contrary to the situation in man, exogenous estrogens cause a fall in serum concentration of the protein.
Animals↗
Characterization of two pregnancy associated serum proteins with pregnancy specific beta 1-glycoprotein (PS-beta-G)-determinants.
The existence of two pregnancy associated, partially identical glycoproteins with PS-beta-G determinants were demonstrated in serum from pregnant women. Crossed and crossed-line immunoelectrophoretic analysis of pregnancy sera revealed two precipitates with alpha- and beta-mobility, respectively. The ratio between the precipitation areas in various samples could be related to three different types of precipitation patterns observed in rocket immunoelectrophoresis. This variation in precipitation pattern could so far not be related to the progression of any obstetric parameters. The diphasic shape of the precipitate in crossed immunoelectrophoresis was not changed by incubation with normal human male serum or antibody to normal human serum. Incorporation of 4% PEG in the gels enhanced the precipitate in the alpha-region. The cross-reactive glycoproteins were detected in all pregnant women examined. These results point to the need for specific antibody preparations permitting separate quantitation of the two antigen populations with PS-beta-G determinants.
Chemical Precipitation↗
Two pregnancy-associated serum proteins with pregnancy-specific beta1-glycoprotein determinants.
A protein showing strong antigenic cross-reactivity with the "pregnancy-specific" beta1-glycoprotein (PSbetaG) was demonstrated in serum of pregnant women during the second and third trimesters, but not in sera from normal men and nonpregnant women. The described protein had alpha2-electrophoretic mobility and its molecular weight was estimated to be around 200,000 as compared to 80,000 for PSbetaG. Crossed immunoelectrophoretic analysis, using antiserum to PSbetaG, indicated the presence of additional antigenic determinants in PSbetaG not demonstrable in the cross-reacting alpha2-protein. The PSbetaG/alpha2-protein ratio was rather constant in consecutive serum samples from one woman during pregnancy, but showed a marked variation when sera from different women were compared. Results of rocket immunoelectrophoresis gave a rough estimate of the ratio between these two proteins in a sample of pregnancy serum. The reported findings point to the need for development of specific methods for quantitation of the two proteins and the necessity to re-evaluate earlier quantitative PSbetaG data.
Blood Protein Electrophoresis↗
Affinity chromatographic purification of the pregnancy zone protein.
An immunospecific affinity chromatographic method for purification of human pregnancy zone protein (PZP) directly from serum is described. Highly purified goat-anti-human PZP-immunoglobulin was applied as a ligand. Recovery of PZP varied from 56--75%. The impurities constituted maximally 5--10% of the total protein in the eluate. The purification factor was approximately 100.
Blood Proteins↗
An immunoprecipitation-dissociation technique for large scale antibody purification and an antigen consumption electroimmunoassay for antibody quantitation. A model study with antibodies to pregnancy zone protein.
A simple immunoprecipitation--dissociation technique for large scale purification of antibodies is described, which comprises selective denaturation of the antigen and recovery of the antibody fraction by exclusion chromatography at low pH. Its use is illustrated by the purification of antibodies to pregnancy zone protein. A purification factor of about 60 was achieved. An antigen consumption electroimmunoassay was also developed which permits quantitative determination of the antigen binding activity of antibodies with a given specificity. The methods have general application.
Antibodies↗
An automatic multi-programmed affinity chromatographic system.
An automatic immunospecific affinity-chromatographic system for continuous operation is described. The system comprises time-controlled sample application, washing and elution steps and automatic dialysis of eluted fractions. The applicability of the system is illustrated by the purification of pregnancy zone protein on immunosorbent gel.
Blood Proteins↗